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Biomedical subjects

A O Hamilton

Publications and source records attributed to A O Hamilton.

5 recordsLinked to original sources

Modulation of monocyte complement synthesis by interferons.

Recombinant Escherichia coli-derived gamma-interferon has been shown to stimulate synthesis of the second component of complement (C2), factor B and C1 inhibitor, but to inhibit synthesis of the third component (C3). alpha- and beta-interferons stimulate synthesis of factor B and C3 inhibitor, inhibit C5 synthesis but do not alter synthesis of C2. alpha- and beta-interferons act synergistically with gamma-interferon to enhance both factor B and C1-inhibitor synthesis.

Cells, Cultured

Complement-subcomponent-C1-inhibitor synthesis by human monocytes.

By using a radioimmunoassay, C1-inhibitor was found to accumulate in the supernatants of human monocyte cultures. The production of this protein was inhibited reversibly by cycloheximide. When C1-inhibitor synthesis was compared with C2 synthesis, it was found that C1-inhibitor synthesis continued, whereas synthesis of C2 appeared to cease after about 7 days in culture. Immunoprecipitation of supernatants of monocyte cultures that had been pulsed with [35S]methionine showed a specific band with an Mr of 105 000. Immunoprecipitates of the lysates revealed a band of Mr 83 000; this was thought to represent a partially or non-glycosylated precursor of C1-inhibitor. C1-inhibitor produced by the monocytes was shown, by using a haemolytic assay, to be functionally active. However, the functional activity of C1-inhibitor was reduced by only 44% in the presence of cycloheximide, whereas the concentration of this protein in cycloheximide-treated culture supernatants fell by more than 93%. This finding suggests that monocytes secrete a second molecule, which inhibits C1 activity but is distinct from classical C1-inhibitor.

Cells, Cultured

Prevention of immune precipitation by purified classical pathway complement components.

The role of the classical pathway of complement in the prevention of immune precipitation has been investigated using purified complement components and immune complexes (IC) consisting of rabbit anti-BSA and BSA. C1 reduced the rate of immune precipitation. As C1q, EDTA treated C1 or C1-inhibitor treated C1 were unable to retard the precipitation of IC, it was concluded that the intact C1 molecule was required for this function. Use of phenylmethylsulphonyl fluoride and benzamidine showed that the enzymatic site on C1 was not required for this activity. C4 and C2 did not affect immune precipitation significantly when C1 was present at the concentrations present in serum. When C3 was added to C1, C4 and C2 precipitation of IC did not occur. These data demonstrate that classical pathway activation alone is sufficient for the prevention of immune precipitation.

Antigen-Antibody Complex

Role of C3 in the control of monocyte C2 production.

Serum-treated antigen-antibody complexes (IC) and DTSP-polymerized C3b and C3c inhibited the production of the second complement component (C2) by monocytes in culture, whereas monomeric C3, C3b, C3bi, C3c or C3d had no effect. The degree of inhibition of dithiobissuccinimidyl propionate (DTSP)-polymerized C3 was proportional to the degree of polymerization, dimer exhibiting less inhibitory activity than larger molecular weight polymers. The inhibitory effect of serum-treated IC and DTSP-polymerized C3b was abrogated by their pretreatment with Fab fragments of anti-C3, or pretreatment of monocytes with Fab fragments of antiserum to the C3b receptor (CR1). We have concluded that the inhibition of C2 production produced by serum-treated IC is due to bound C3b or C3bi, and is mediated by CR1. Furthermore cross linking of receptors is required for this effect; two or more than two receptors must be cross-linked for a significant effect to be observed.

Antigen-Antibody Complex

Serum-treated antigen-antibody complexes inhibit the production of C2 and factor B by mononuclear phagocytes.

Antigen-antibody complexes enhanced the synthesis of C2 and factor B by human monocytes and macrophages, and C2 by guinea-pig macrophages. In contrast complexes that had been treated with serum inhibited the production of these components. The inhibitory effect of serum-treated complexes was abrogated by Fab fragments of anti-C3, anti-C3c and anti-C3d. It is therefore probable that inhibition was mediated by a C3 fragment bound to the complex. The enhancing effect of untreated complexes was reversible by serum-treated complexes, and the inhibitory action of serum-treated complexes was counteracted by untreated complexes. Such a system may be important in the regulation of the synthesis of complement components in response to local requirements.

Animals